1. MinD tethers DNA to liposomes in an ATP‐dependent manner.
MinD tethers DNA to liposomes in an ATP‐dependent manner. (A) Schematic overview of the flotation experiment with DOPG (1,2‐dioleoyl‐sn‐glycero‐3‐phospho‐(1′‐rac‐glycerol)) liposomes, fluorescently labeled with 0.1% DiO (3,3′‐dioctadecyloxacarbocyanine perchlorate). Upon ultracentrifugation in the Nycodenz density gradient, free liposomes move to the top of the gradient (fraction 1), whereas soluble material remains at the bottom (fractions 3 and 4). Heavier (high density) liposome–protein complexes are found in fraction 2. Relative distribution of DiO‐labeled DOPG liposomes (400 μg/ml) to individual fractions in the flotation assay in the absence (B) or presence (C) of MinD (1.3 μM). (D) Flotation assays with MinD mutants, performed as in (C) including HEX‐labeled DNA (155 bp, P1 promoter, 200 fmol) to the reactions. Only fraction 2 is shown here; other fractions are shown in Supplementary Figure S3D–G. 1 mM ADP or ATP was added to the reaction, as indicated. The x axes represent the amount of material that floted to each fraction as the percentage of the total material recovered from the gradient. (E) Bar plot showing the ratio between the amounts of DNA and DOPG liposomes found in fractions 1 and 2. For the DOPG+DNA case, only fraction 1 is shown, as liposomes do not flot to fraction 2 in the absence of MinD. (F) Bar plot showing the ratio between the amounts of DNA and MinD found in fractions 1 and 2.
Barbara Di Ventura et al. Mol Syst Biol 2013;9:686
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